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Shih JC, Rho J. 
“The Specific Interaction Between LSD and Serotonin-Binding Protein”. 
Res.Commun.Chem.Pathol.Pharmacol.. 1977;16(4):637-47.
Abstract
The chemical interaction between LSD and serotonin-binding protein (SBP) was studied. Methods SBP's were isolated using serotonin affinity chromatography as previously described; rat hypothalamic synaptosomal membrane was passed through a serotonin-Sepharose column. The protein was eluted by to 4 M LSD or to 4 M chlorimipramine (CP) Using a 3-dimensional spectroscopic technique, the fluorogram of free LSD and SBP-bound LSD were determined, from a 0.3 ml aliquot. Ihe specificity of LSD binding was tested by incubating SBP protein or synaptosomal membrane fragments with to , LSD in 0.5 mM Tris pH 7.6 buffer with i% ascorbic acid at 37¡ for to min. After centrifugation at 30,000 x8, the supernatant of this reaction mixture was analyzed fluorometrically. Bovine serum albumin (2 mg BSA) was used as control for the specificity determination. 2 SBP's were isolated. The LSD-eluted SBP produced 2 fluorescence peaks, 1 at 330 am and the other at 465 nm. Peak I (excitation maximum at 280 nm) represents the intrinsic fluorescence of receptor protein, peak 2 (excitation maximum at 375 nm) delineates the fluorescence property of SBP-bound LSD. This LSD bound tightly to SBP and was not dialyzable. LSD alone produced a fluorescence maximum at 435 nm and an excitation maximum at 330 nm. SBP and synaptosomal membrane fragments produced similar fluorograms in the specificity test, but BSALSD mixture produced 2 fluorescence peaks. Peak 1 was a typical fluorescence peak for protein at 340 nm; peak 2 was the fluorescence peak of LSD at 435 nm. similar to that of free LSD. The striking shift of the fluorescence spectra of the SBP-bound LSD, absent when LSD is added to BSA, suggests specific binding between LSD and SBP occurs. After incubation with SBP, the CP fluorescence peak also shifted to considerably longer wavelength regions. The drug-protein interaction caused extensive delocalization of the molecular orbital electrons, lengthening the electronic conjugation of the drug molecule. The convenience of the 3 dimensional spectroscopic technique is stressed.
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